Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

The bacterial virulence factor InlC perturbs apical cell junctions and promotes cell-to-cell spread of Listeria


Several pathogenic bacteria, including Listeria monocytogenes, use an F-actin motility process to spread between mammalian cells1. Actin 'comet tails' propel Listeria through the cytoplasm, resulting in bacteria-containing membrane protrusions that are internalized by neighbouring cells. The mechanism by which Listeria overcomes cortical tension to generate protrusions is unknown. Here, we identify bacterial and host proteins that directly regulate protrusions. We show that efficient spreading between polarized epithelial cells requires the secreted Listeria virulence protein InlC (internalin C). We next identify the mammalian adaptor protein Tuba as a ligand of InlC. InlC binds to a carboxy-terminal SH3 domain in Tuba, which normally engages the human actin regulatory protein N-WASP2. InlC promotes protrusion formation by inhibiting Tuba and N-WASP activity, probably by impairing binding of N-WASP to the Tuba SH3 domain. Tuba and N-WASP are known to control the structure of apical junctions in epithelial cells3. We demonstrate that, by inhibiting Tuba and N-WASP, InlC makes taut apical junctions become slack. Experiments with myosin II inhibitors indicate that InlC-mediated perturbation of apical junctions accounts for the role of this bacterial protein in protrusion formation. Collectively, our results suggest that InlC promotes bacterial dissemination by relieving cortical tension, thereby enhancing the ability of motile bacteria to deform the plasma membrane into protrusions.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Figure 1: InlC is needed for efficient spreading and protrusion formation in a polarized cell line.
Figure 2: InlC interacts with the mammalian adaptor protein Tuba.
Figure 3: Tuba and N-WASP control protrusion formation.
Figure 4: InlC, Tuba and N-WASP control the morphology of apical junctions.
Figure 5: Model for InlC-mediated cell-to-cell spread of Listeria.

Similar content being viewed by others


  1. Gouin, E., Welch, W. D. & Cossart, P. Actin-based motility of intracellular pathogens. Curr. Opin. Microbiol. 8, 35–45 (2005).

    Article  CAS  Google Scholar 

  2. Miki, H. & Takenawa, T. Regulation of actin dynamics by WASP family proteins. J. Biochem. 134, 309–313 (2003).

    Article  CAS  Google Scholar 

  3. Otani, T., Ichii, T., Aono, S. & Takeichi, M. Cdc42 GEF Tuba regulates the junctional configuration of simple epithelial cells. J. Cell Biol. 175, 135–46 (2006).

    Article  CAS  Google Scholar 

  4. Vazquez-Boland, J. A. et al. Listeria pathogenesis and molecular virulence determinants. Clin. Microbiol. Rev. 14, 584–640 (2001).

    Article  CAS  Google Scholar 

  5. Robbins, J. R. et al. Listeria monocytogenes exploits normal host cell processes to spread from cell to cell. J. Cell Biol. 146, 1333–1349 (1999).

    Article  CAS  Google Scholar 

  6. Monack, D. M. & Theriot, J. A. Actin-based motility is sufficient for bacterial membrane protrusion formation and host cell uptake. Cell. Microbiol. 3, 633–647 (2001).

    Article  CAS  Google Scholar 

  7. Miyoshi, J. & Takai, Y. Molecular perspective on tight-junction assembly and epithelial polarity. Adv. Drug Deliv. Rev. 57, 815–855 (2005).

    Article  CAS  Google Scholar 

  8. Lecuit, M. et al. A transgenic model for Listeriosis: role of internalin in crossing the intestinal barrier. Science 292, 1722–1725 (2001).

    Article  CAS  Google Scholar 

  9. Peterson, M. D. & Mooseker, M. S. Characterization of the enterocyte-like brush border cytoskeleton. J. Cell. Sci. 102, 581–600 (1992).

    CAS  PubMed  Google Scholar 

  10. Sun, A. N., Camilli, A. & Portnoy, D. A. Isolation of Listeria monocytogenes small-plaque mutants defective for intracellular growth and cell-to-cell spread. Infect. Immun. 58, 3770–3778 (1990).

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Engelbrecht. F. et al. A new PrfA-regulated gene of Listeria monocytogenes encoding a small, secreted protein which belongs to the family of internalins. Mol. Microbiol. 21, 823–837 (1996).

    Article  CAS  Google Scholar 

  12. Smith, G. A. et al. The two distinct phospholipases C of Listeria monocytogenes have ovelapping roles in escape from a vacuole and cell-to-cell spread. Infec. Immun. 63, 4231–4237 (1995).

    CAS  Google Scholar 

  13. Smith, G. A., Theriot, J. & Portnoy, D. The tandem repeat domain in the Listeria monocytogenes ActA protein controls the rate of actin-based motility, the percentage of moving bacteria, and the localization of vasodilator-stimulated phosphoprotein and profilin. J. Cell Biol. 135, 647–660 (1996).

    Article  CAS  Google Scholar 

  14. Fievet, B., Louvard, D. & Arpin, M. ERM proteins in epithelial cell organization and functions. Biochim. Biophys. Acta. 1773, 653–660 (2007).

    Article  CAS  Google Scholar 

  15. Pust, S., Morrison, H., Wehland, J., Sechi, A. S. & Herrlich, P. Listeria monocytogenes exploits ERM protein functions to efficiently spread from cell to cell. EMBO J. 24, 1287–1300 (2005).

    Article  CAS  Google Scholar 

  16. Sechi, A. S., Wehland, J. & Small, J. V. The isolated comet tail pseudopodium of Listeria monocytogenes: a tail of two actin filament populations, long and axial and short and random. J. Cell Biol. 137, 155–167 (1997).

    Article  CAS  Google Scholar 

  17. Kobe, B. & Kajava, A. V. The leucine-rich repeat as a protein recognition motif. Curr. Opin. Struct. Biol. 11, 725–732 (2001).

    Article  CAS  Google Scholar 

  18. Bierne, H., Sabet, C., Personnic, N. & Cossart, P. Internalins: a complex family of leucine-rich repeat-containing proteins in Listeria monocytogenes. Microbes Infec. 9, 1156–66 (2007).

    Article  CAS  Google Scholar 

  19. Salazar, M. A. et al. Tuba, a novel protein containing Bin-Amphysin-Rvs and Dbl homology domains, links Dynamin to regulation of the actin cytoskeleton. J. Biol. Chem. 278, 49031–49043 (2003).

    Article  CAS  Google Scholar 

  20. Kovacs, E. M., Makar, R. S. & Gertler, F. B. Tuba stimulates N-WASP-dependent actin assembly. J. Cell Sci. 119, 2715–2726 (2006).

    Article  CAS  Google Scholar 

  21. Li, S. S. C. Specificity and versatility of SH3 and other proline-recognition domains: structural basis and implications for cellular signal transduction. Biochem. J. 390, 641–653 (2005).

    Article  CAS  Google Scholar 

  22. Straight, A. F. et al. Dissecting temporal and spatial control of cytokinesis with a myosin II inhibitor. Science 299, 1743–1747 (2003).

    Article  CAS  Google Scholar 

  23. Riento, K. & Ridley, A. J. Rocks: multifunctional kinases in cell behaviour. Nature Rev. Mol. Cell Biol. 4, 456–456 (2003).

    Article  Google Scholar 

  24. Nusrat, A., Turner, J. R. & Madara, J. L. Molecular physiology and pathophysiology of tight junctions IV. Regulation of tight junctions by extracellular stimuli: nutrients, cytokines, and immune cells. Am. J. Physiol. Gastrointest. Liver Physiol. 279, G851–G857 (2000).

    Article  CAS  Google Scholar 

  25. Wuenscher, M. D., Kohler, S., Goebel, W. & Chakraborty, T. Gene disruption by plasmid integration in Listeria monocytogenes: insertional inactivation of the listeriolysin determinant lisA. Mol. Gen. Genet. 228, 177–182 (1991).

    Article  CAS  Google Scholar 

  26. Co, C., Wong, D. T., Gierke, S., Chang, V. & Taunton, J. Mechanism of actin network attachment to moving membranes: barbed end capture by N-WASP WH2 domains. Cell 128, 901–913 (2007).

    Article  CAS  Google Scholar 

  27. Sun, H. et al. Host adaptor proteins Gab1 and CrkII promote InlB-dependent entry of Listeria monocytogenes. Cell. Microbiol. 7, 443–457 (2005).

    Article  CAS  Google Scholar 

  28. Auerbuch, V., Loureiro, J. J., Gertler, F. B., Theriot, J. A. & Portnoy, D. A. Ena-VASP proteins contribute to Listeria monocytogenes pathogenesis by controlling temporal and spatial persistence of bacterial actin-based motility. Mol. Microbiol. 49, 1361–1375 (2003).

    Article  CAS  Google Scholar 

Download references


We thank J. Brumell, H. Sarantis, A. Wilde and R. Collins for reviewing the manuscript, N. Freitag, T. Otani, E. Leung and K. Nemec for advice on assays or protein purification and Y. Shen for help with plasmid construction. This work was supported by grants from the Canadian Institutes of Health Research (CIHR) (MT-15497) and National Institutes of Health (1R21AI076881-01) to K.I., and a CIHR grant (MOP-15499) to S.D.G.

Author information

Authors and Affiliations



K.I., T.R. and S.D.G.O. designed research; T.R. and B.G. performed the research; T.R. and K.I. analysed data; M.H., S.M.A. and W.G. contributed novel reagents and K.I. and T.R. wrote the paper.

Corresponding author

Correspondence to Keith Ireton.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary Information (PDF 790 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rajabian, T., Gavicherla, B., Heisig, M. et al. The bacterial virulence factor InlC perturbs apical cell junctions and promotes cell-to-cell spread of Listeria. Nat Cell Biol 11, 1212–1218 (2009).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

This article is cited by


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing